Klezovitch O, Edelstein C, Scanu A M
Department of Medicine, University of Chicago, Illinois 60637, USA.
J Clin Invest. 1996 Jul 1;98(1):185-91. doi: 10.1172/JCI118765.
It is now established that the lysine binding site (LBS) of apo(a) kringle IV-10, and particularly Trp72, plays a dominant role in the binding of lipoprotein(a) [Lp(a)] to lysine. To determine the role of the LBS in the binding of Lp(a) to fibrinogen, we examined the binding to plasmin-modified (PM) fibrinogen of human and rhesus monkey Lp(a) species classified as either Lys' or Lys- based on their capacity to bind lysine Sepharose and to have Trp or Arg, respectively, in position 72 of the LBS of kringle IV-10. We also examined the free apo(a)s obtained by subjecting their corresponding parent Lp(a)s to a mild reductive procedure developed in our laboratory. Our results show that both Lyst and Lys- Lp(a)s and their derived apo(a)s, bound to PM-fibrinogen with similar affinities (Kds: 33-100 nM), whereas the B(max) values were threefold higher for apo(a)s. Both the lysine analog epsilon-aminocaproic acid and L-proline inhibited the binding of Lp(a) and apo(a) to PM fibrinogen. We conclude that the LBS of kringle IV-10 is not involved in this process and that apo(a) binds to PM-fibrinogen via a lysine-proline-sensitive domain located outside the LBS and largely masked by the interaction of apo(a) with apoB100. The significant difference in the PM fibrinogen binding capacity also suggests that apo(a) may have a comparatively higher athero-thrombogenic potential than parent Lp(a).
现已确定,载脂蛋白(a) kringle IV-10的赖氨酸结合位点(LBS),尤其是色氨酸72,在脂蛋白(a) [Lp(a)]与赖氨酸的结合中起主导作用。为了确定LBS在Lp(a)与纤维蛋白原结合中的作用,我们检测了根据其结合赖氨酸琼脂糖的能力以及kringle IV-10的LBS第72位分别具有色氨酸或精氨酸而分类为Lys⁺或Lys⁻的人和恒河猴Lp(a)物种与纤溶酶修饰(PM)纤维蛋白原的结合情况。我们还检测了通过对其相应的亲本Lp(a)进行我们实验室开发的温和还原程序而获得的游离载脂蛋白(a)。我们的结果表明,Lys⁺和Lys⁻ Lp(a)及其衍生的载脂蛋白(a)以相似的亲和力(解离常数:33 - 100 nM)与PM纤维蛋白原结合,而载脂蛋白(a)的最大结合量(Bmax)值高3倍。赖氨酸类似物ε-氨基己酸和L-脯氨酸均抑制Lp(a)和载脂蛋白(a)与PM纤维蛋白原结合。我们得出结论,kringle IV-10的LBS不参与此过程,并且载脂蛋白(a)通过位于LBS之外且在很大程度上被载脂蛋白(a)与载脂蛋白B100的相互作用所掩盖的赖氨酸-脯氨酸敏感结构域与PM纤维蛋白原结合。PM纤维蛋白原结合能力的显著差异还表明,载脂蛋白(a)可能比亲本Lp(a)具有相对更高的动脉粥样硬化血栓形成潜力。